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6 protocols using artemether

1

Lateral Flow Assay for Antimalarial Drugs

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Carboxylate-modified TRFN, 1% solids, 200 nm, 580/605 (λexem), and CGN, 30 nm, were purchased from Shanghai Jieyi Biotechnology. Three haptens of artemether, dihydroartemisinin, and artesunate conjugated with bovine serum albumin (BSA), three monoclonal antibodies (mAbs) against artemether, dihydroartemisinin, and artesunate were produced in our laboratory [11 (link), 12 (link), 15 (link)].
1-Ethyl-3-(3-(dimethylamino) propyl) carbodiimide hydrochloride (EDC) and goat anti-mouse IgG were purchased from Sigma (St. Louis, MO, USA). Artemether, dihydroartemisinin and artesunate standards were purchased from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). The nitrocellulose filter membrane (NC membrane), sample pad, conjugate pad, absorbance pad, support plate, the portable optical scanner, and the fluorescence lateral flow reader were obtained from Shanghai Jinbiao Biotechnology Co., Ltd. (Shanghai, China). The other chemicals and organic solvents used were of analytical grade and purchased from Sinopharm Chemical Reagent (Beijing, China). The commercial antimalarial drugs used for detection were collected from Africa and Southeast Asia.
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2

Anti-malarial Compounds Characterization

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DSM267 and DSM265 were gifts from M. Phillips from University of Texas (UT) Southwestern (35 (link)). l-dihydroorotic acid (DHO) was purchased from Sigma-Aldrich (St. Louis, MO). Amodiaquine, artemether, atovaquone, chloroquine, dihydroartemisinin, and mefloquine were purchased from Sigma-Aldrich (St. Louis, MO).
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3

Neuroblastoma Cell Line Transfection

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Human neuroblastoma cell lines SK-N-SH, SH-SY5Y, and SK-N-BE2 were purchased from the American Type Culture Collection (Manassas, VA, USA). The cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) in an atmosphere containing 5% CO2 at 37°C. Artemether and doxorubicin were purchased from Sigma-Aldrich (St. Louis, MO, USA). For transfection, neuroblastoma cells were transiently transfected with the recombinant plasmid pcDNA(−)3.1-B7-H3 or control pcDNA(−)3.1. At 48 h after transfection, the transfection efficiency was evaluated by real-time PCR and Western blot analysis.
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4

Artemether Dilution and Storage

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Artemether (80 mg·mL−1) were obtained from Sigma‐Aldrich Co. (St Louis, MO, USA) and dissolved in 80 mL of sterile water to form 10 mg·mL−1 storage solution conserved at 4 °C and then diluted to the required concentration with DMEM (4500 mg·L−1 glucose, 10% fetal bovine serum, 1% penicillin/streptomycin) when used.
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5

Artemether and Sulfadiazine Cytotoxicity Assay

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Artemether (methyl-ether-qinghaosu) was purchased from Exim Pharm Co. (USA). Artemether was prepared in ethanol–water (v/v) (30–70%) with 1000 µg/ml concentration, then 5, 10, 25, 50, and 100 µg/ml dilutions were made from stock solutions in DMEM.[18 ] Sulfadiazine was obtained from Sigma (Sigma-Aldrich) and solved in DMSO (dimethyl sulfoxide) as stock solutions with 1000 µg/ml concentration. Following 1.6, 3.12, 6.25, 12.5, 25, 50, 100, and 200 µg/ml dilutions were prepared in DMEM. Provided solutions were stored at 4°C and used in cytotoxicity assays and in vitro experiments.[1 (link)]
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6

Artemether Modulation of LPS Response

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Artemether (Sigma) was dissolved in dimethyl sulphoxide (DMSO) to a concentration of 0.1 M and aliquots stored in -80°C. BAY 11-7085 and SKF 86002 dihydrochloride were purchased from Tocris. In all experiments, cells were treated with Artemether
(5-40 µM) in the absence or presence of LPS (1 µg/ml).
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